大豆脂肪酶SDP1生物信息学分析和基因编辑
- Title:
- Bioinformatics Analysis and Gene Editing of Soybean SDP1 Lipase
- 关键词:
- 大豆; SDP1基因; CRISPR/Cas9技术; 生物信息学分析; 组织特异性表达
- 摘要:
- 三酰甘油脂肪酶（Suger-dependent1，SDP1）是一种脂解酶，在植物发育过程中发挥重要作用。为发掘大豆SDP1基因的生物学功能，利用生物信息学分析方法对大豆GmSDP1基因的进化关系、保守基序和启动子区域的顺式作用元件等进行分析。利用基因编辑技术创制大豆gmsdp1-1CR突变体，对其编辑事件、含油量以及脂肪酸含量进行分析。结果表明：4个大豆GmSDP1基因与双子叶植物亲缘关系较近，保守结构域和蛋白二级结构高度相似，启动子区域含有抗逆境胁迫以及激素相关的调控元件。对GmSDP1在不同组织部位的表达量分析发现，GmSDP1在子叶、叶片和花等部位表达较高。运用CRISPR-Cas9基因编辑技术创制大豆Willimas 82品种的gmsdp1-1CR突变体，设计1条靶向目的基因GmSDP1-1的特异靶点，构建pGES201-GmSDP1-1敲除载体。毛根转化试验的阳性大豆毛状根编辑效率达到56.52%。大豆稳定转化的T1代基因编辑阳性植株编辑效率为1.5%，选取gmsdp1-1CR突变体做进一步表型分析表明，油酸含量极显著降低，亚麻酸含量极显著升高。研究结果为深入研究GmSDP1基因的功能和调控机制提供了参考依据。
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